The present invention relates to a pharmaceutical composition having antirheumatic activity, antithrombotic activity, analgetic activity, antipyretic activity, anti-hyperlipemic activity and anti-inflammatory activity and activities of reducing the level of blood sugar, raising the coronary blood flow, improving the capability of deformation of erythrocytes, reducing the blood pressure, ameliorating proteinuria and proteinemia and regulating production or metabolism of prostaglandins in a mammal, containing a glycoprotein derived from a basidiomycetous fungus belonging to the genus Coriolus, for instance, Coriolus versicolor (Fr.) Quel., as an active ingredient. The glycoprotein derived from Coriolus versicolor (Fr.) Quel. [FERM-P No. 2412 (ATCC 20547)] has been already supplied to the public as an anti-tumour drug under the trade mark of Krestin.
Since the glycoprotein is low in mammalian toxicity and does not disturb the intestinal microflora, the pharmaceutical composition containing the glycoprotein as an active ingredient can be administered for a long time period. In addition, the glycoprotein is quite free of the fear of causing malformation and/or allergic reaction and accordingly, the glycoprotein is an extremely safe substance.
The active ingredient, the glycoprotein, of the pharmaceutical composition according to the present invention is a publicly known substance, and as has been disclosed in Japanese Patent publications Nos. 17149/1971, 36322/1976, 14274/1981, 14276/1981 and 39288/1981, the glycoprotein is obtained by culturing a basidiomycetous fungal species belonging to the genus Coriolus, extracting the thus proliferated mycelia or fruit bodies with hot water or an aqueous alkali solution, and removing low molecular weight substances having a molecular weight of less than 5000 and the thus obtained substance as an extract contains from about 18 to 38% by weight of proteins and shows a molecular weight of from 5,000 to 300,000 as determined by ultracentrifugal method.
The glycoprotein derived from the mycelia of Coriolus versicolor (Fr.) Quel. is liver brown in color and has a nitrogen content of 2 to 8%, in many cases 3 to 6%. Various color reaction tests on the glycoprotein according to the present invention gave the results as shown below.
______________________________________ naphthol sulfuric acid reaction Purple (Molish's reaction) Indole sulfuric acid reaction Brown (Dische's reaction) Anthrone sulfuric acid reaction Greenish blue Phenol sulfuric acid reaction Brown Tryptophane sulfuric acid reaction Purplish brown Lowry-Folin process Blue Ninhydrin reaction after hydrochloric Greenish blue acid hydrolysis ______________________________________
The molecular weight of the glycoprotein according to the present invention is 5000 to 300,000 as measured according to an ultracentrifugal method. The glycoprotein according to the present invention contains about 18 to 38% by weight of proteins.
The saccharide moiety of the glycoprotein of the present invention consists mainly of .beta.-D-glycan and the structure of the glycan moiety is a branched one containing 1.fwdarw.3, 1.fwdarw.4 and 1.fwdarw.6 bondings. Of the amino acids forming the protein moiety of the glycoprotein, the amount of acidic amino acids such as aspartic acid, glutamic acid, etc. and that of neutral amino acid such as valine, leucine, etc. are relatively large, and the amount of basic amino acids such as lysine, arginine, etc. is relatively small. The glycoprotein is soluble in water and almost insoluble in hexane, benzene, chloroform, methanol and pyridine. The glycoprotein slowly decomposes at a temperature around 120.degree. C. when it is heated.
As will be seen in Table 1, the mammalian toxicity of the glycoprotein of the present invention is extremely low, and it hardly causes any side effects on animals. Namely, it is known to be a very safe substance to living bodies.
TABLE 1 ______________________________________ Animal Route of LD.sub.50 (mg/kg) species administration Female Male ______________________________________ Mouse intravenous &gt;1300 &gt;1300 Strain subcutaneous &gt;5000 &gt;5000 ICR-JCL intraperitoneal &gt;5000 &gt;5000 oral &gt;20000 &gt;20000 Rat intravenous &gt;600 &gt;600 Strain subcutaneous &gt;5000 &gt;5000 Donryu intraperitoneal &gt;5000 &gt;5000 oral &gt;20000 &gt;20000 ______________________________________
The mice used in the test for finding the above-mentioned acute toxicity value (LD.sub.50 mg/kg) were of the strain ICR-JCL, 4 to 5 weeks after birth, and body weight of 21 to 24 g. The rats used in the same test were of the strain Donryu, 4 to 5 weeks after birth, and body weight of 100 to 150 g. The glycoprotein was dissolved in a physiological saline and administered via each route shown in Table 1. After administration, observation on the general symptoms, mortality and body weight of each of the thus treated animals were carried out for 7 days, and then they were sacrificed to be subjected to autopsy.
As are shown in Table 1, no case of death was found both on the mice and the rats even at the maximum dosage which could be given and accordingly, the glycoprotein of the present invention is extremely safe for living body to the extent that the value of LD.sub.50 could not be actually determined.
As a result of examining the physiological and pharmaceutical properties of the glycoprotein derived from a basidiomycetous fungus belonging to the genus Coriolus, it has been found out that the glycoprotein shows antirheumatic activity, antithrombotic activity, analgetic activity, antipyretic activity, anti-hyperlipemic activity and anti-inflammatory activity and activities of reducing the level of blood sugar, raising the coronary blood flow, improving the capability of deformation of erythrocytes, reducing the blood pressure, ameliorating proteinuria and proteinemia and regulating production or metabolism of prostaglandins in a mammal, as well as the anti-tumour activity, and based on the findings, the present inventors have attained the present invention.